• Title/Summary/Keyword: Cellular association

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Molecular Characterization and Expression Analysis of Insulin-like Growth Factor-1 and Insulin-like Growth Factor Binding Protein-1 Genes in Qinghai-Tibet Plateau Bos grunniens and Lowland Bos taurus

  • Chen, Ya-bing;Fu, Mei;Lan, Dao-liang;Li, Jian
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.1
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    • pp.20-24
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    • 2015
  • Insulin-like growth factor-1 (IGF-1) and insulin-like growth factor binding protein-1 (IGFBP-1) play a pivotal role in regulating cellular hypoxic response. In this study, we cloned and characterized the genes encoding IGF-1 and IGFBP-1 to improve the current knowledge on their roles in highland Bos grunniens (Yak). We also compared their expression levels in the liver and kidney tissues between yaks and lowland cattle. We obtained full-length 465 bp IGF-1 and 792 bp IGFBP-1, encoding 154 amino acids (AA) IGF-1, and 263 AA IGFBP-1 protein, respectively using reverse transcriptase-polyerase chain reaction (RT-PCR) technology. Analysis of their corresponding amino acid sequences showed a high identity between B. grunniens and lowland mammals. Moreover, the two genes were proved to be widely distributed in the examined tissues through expression pattern analysis. Real-time PCR results revealed that IGF-1 expression was higher in the liver and kidney tissues in B. grunniens than in Bos taurus (p<0.05). The IGFBP-1 gene was expressed at a higher level in the liver (p<0.05) of B. taurus than B. grunniens, but it has a similar expression level in the kidneys of the two species. These results indicated that upregulated IGF-1 and downregulated IGFBP-1 are associated with hypoxia adaptive response in B. grunniens.

Molecular Cloning, Tissue Distribution and Expression of Porcine y+L Amino Acid Transporter-1

  • Zhi, Ai-min;Zhou, Xiang-yan;Zuo, Jian-jun;Zou, Shi-geng;Huang, Zhi-yi;Wang, Xiao-lan;Tao, Lin;Feng, Ding-yuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.2
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    • pp.272-278
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    • 2010
  • In this study, we cloned, sequenced and characterized porcine y+L Amino Acid Transporter-1 (y+LAT1). By screening a translated EST database with the protein sequence of the human $y^{+}$LAT1 and by using rapid amplification of cDNA ends (RACE), the full-length cDNA encoding porcine $y^{+}$LAT1 was isolated from porcine intestine RNA. It was 2,111 bp long, encoding a 511 amino acid trans-membrane glycoprotein composed of 12 transmembrane domains. The predicted amino acid sequence was found to be 91%, 90%, 87% and 87% identical to those of cattle, human, mouse and rat $y^{+}$LAT1 respectively. Real-time RT-PCR results indicated that the small intestine had the highest $y^{+}$LAT1 mRNA abundance and the lung had the lowest $y^{+}$LAT1 mRNA abundance. Baby hamster kidney (BHK) cells transfected with green fluorescent protein (GFP) tagged porcine $y^{+}$LAT1 cDNA indicated that the cellular localization of the gene product in BHK was on the plasma membrane.

Establishment and Characterization of the Fibroblast Line from Silkie Bantam

  • Li, L.F.;Guan, W.J.;Li, H.;Bai, X.J.;Ma, Y.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.4
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    • pp.492-499
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    • 2009
  • A Silkie Bantam embryo fibroblast line (named SBF59 line) was successfully established by using direct explant culture and cryopreservation techniques. Cell morphology, viability, dynamic growth and contamination were tested and the karyotype and levels of isoenzymes of lactic dehydrogenase and malic dehydrogenase were analyzed. Four kinds of fluorescent protein extrogenes, including $pEGFP-N_3$, $pECFP-N_1$, $pEYFP-N_1$ and $pDsRed1-N_1$ were transfected into the cells. The results showed that the cells were healthy and possessed a fibrous structure without a change in morphology. The average viability of the cells was 96% before freezing and 90.5% after thawing. The growth curve appeared as typical "S" shape and the cell growth passed through a detention phase, a logarithmic phase and a platform phase; the estimated population doubling time (PDT) was 38.5 h; assays for the presence of bacteria, fungi, viruses and mycoplasmas were negative; the cell line showed no cross contamination when assessed by isoenzyme analysis; the chromosome number was 2n = 78 on more than 88% of occasions; four kinds of fluorescent protein extro-genes appeared to be expressed effectively with a high transfection efficiency between 18.3% and 42.3%. The cell line met the required quality control standard. It not only preserves the genetic resources of the important Silkie Bantam at the cellular level but also provides valuable materials for genomic, post-genomic, somatic cell cloning research and other applications.

Variations in mitochondrial cytochrome b region among Ethiopian indigenous cattle populations assert Bos taurus maternal origin and historical dynamics

  • Tarekegn, Getinet Mekuriaw;Ji, Xiao-yang;Bai, Xue;Liu, Bin;Zhang, Wenguang;Birungi, Josephine;Djikeng, Appolinaire;Tesfaye, Kassahun
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.9
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    • pp.1393-1400
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    • 2018
  • Objective: This study was carried out to assess the haplotype diversity and population dynamics in cattle populations of Ethiopia. Methods: We sequenced the complete mitochondrial cytochrome b gene of 76 animals from five indigenous and one Holstein Friesian${\times}$Barka cross bred cattle populations. Results: In the sequence analysis, 18 haplotypes were generated from 18 segregating sites and the average haplotype and nucleotide diversities were $0.7540{\pm}0.043$ and $0.0010{\pm}0.000$, respectively. The population differentiation analysis shows a weak population structure (4.55%) among the populations studied. Majority of the variation (95.45%) is observed by within populations. The overall average pair-wise distance ($F_{ST}$) was 0.049539 with the highest ($F_{ST}=0.1245$) and the lowest ($F_{ST}=0.011$) $F_{ST}$ distances observed between Boran and Abigar, and Sheko and Abigar from the indigenous cattle, respectively. The phylogenetic network analysis revealed that all the haplotypes detected clustered together with the Bos taurus cattle and converged to a haplogroup. No haplotype in Ethiopian cattle was observed clustered with the reference Bos indicus group. The mismatch distribution analysis indicates a single population expansion event among the cattle populations. Conclusion: Overall, high haplotype variability was observed among Ethiopian cattle populations and they share a common ancestor with Bos taurus.

Association Between Survivin Gene Polymorphisms and the Susceptibility to Colon Cancer Development in the Turkish Population

  • Yamak, Nesibe;Yaykasli, Kursat Oguz;Yilmaz, Umit;Eroz, Recep;Uzunlar, Ali Kemal;Ankarali, Handan;Sahiner, Cem;Baltaci, Davut
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.8963-8967
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    • 2014
  • Background: Colon cancer is one of the most common cancers worldwide. Apoptosis is a necessary physiological process for cell elimination which is very important both cellular homeostasis and cell proliferation and differantiation. Dysregulation can lead to uncontrolled cell growth and tumor development. Survivin, a member of the IAP family, plays a key role in promotion of cell proliferation as well as inhibition of apoptosis in cancer cells. The aim of this study was to investigate whether specific genetic polymorphisms of survivin could be associated with colon cancer development and progression in a Turkish population. Our study is the first to our knowledge to investigate the relationship between colon cancer risk and survivin gene polymorphisms. Materials and Methods: The relation between colon cancer and survivin -31 G/C (rs9904341), -241 C/T (rs17878467) and -625 C/G (rs8073069) polymorphism in promotor site of survivin gene associated with apoptosis was investigated using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Results: Individuals with -31C allele and CC genotype were found to have a higher risk of developing colon cancer (OR=13.4, p=0.01). The -241 CT genotype considerably increased the risk of colon cancer (OR=12.0, p=0.0001). However, there was no significant varaition of the survivin -625 C/G polymorphism among colon cancer patients and controls in our study. Conclusions: This study provides the first evidence that survivin -31 G/C and -241 C/T SNP significantly contribute to the risk of colon cancer in the Turkish population.

Production of Monoclonal Antibodies (Mabs) Against Surface Antigens on Israeli Carp Lymphocytes and Their Applications

  • Woo, Jong-Kyu;Jang, Han-Na;Cho, Young-Hye;Jang, Yong-Suk;Choi, Sang-Hoon
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.8
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    • pp.1179-1187
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    • 2001
  • In fish both humoral and cell mediated immune responses have been reported whereas antibodies recognizing specific cellular populations have not yet been developed except for ones recognizing surface Ig molecules on B lymphocytes. Our aim was to develop and characterize monoclonal antibodies (Mabs) specific for the immune-related cells. Mabs were produced by fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells previously sensitized against Israeli carp (I. carp) kidney mononuclear cells. We obtained 44 Mabs positively reacting with I. carp kidney mononuclear cells and partially characterized 7 Mabs in the morphological and mitogen-based proliferative aspects. Fluorescence-activated cell sorter (FACS) analysis against I. carp kidney cells by using 7 different Mabs showed 80.3% for ICK 17-4, 65.1% for ICK 2-3, 64.1% for ICK 25-1, 67.5% for lCK 22-1, 70.8% for ICK 16-2, 76.8% for ICK 13-2, 79.7% for ICK II-I. Panning method was used for the isolation of Mabs specific mononuclear carp spleen cells followed by Wright's stain. The stained cell populations were identified as monocytes (ICK 17-4, ICK 2-3, ICK 25-1, ICK 22-1 and ICK 16-2), lymphocytes (ICK 11-1), and a mixed cell population of monocytes and lymphocytes (ICK 13-2). In cell proliferation assay, monocytes purified by ICK 17-4, 2-3 and 22-1 efficiently responded to Con A and PHA, while ones separated by ICK 25-1 did not react with any mitogens. Lymphocytes isolated by ICK 11-1, though it is not known whether they are T or B cells, were more responsive to Con A than PHA or LPS, suggesting that fish immune cells are somewhat different from mammalian cells in responding to mammalian T or B cell mitogens.

The therapeutic effects of WSY-0702 on benign prostatic hyperplasia in RWPE-1

  • Oh, Hyun-A;Kwon, Eun Bi;Hwang, Yun Gyeong;Park, Soon Eung;Mok, Ji Ye;Hwang, Sung Yeoun
    • CELLMED
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    • v.7 no.2
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    • pp.8.1-8.7
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    • 2017
  • Benign prostatic hyperplasia (BPH) is one of the major diseases of the urinary system in older men. WSY-0702 is the extracted from the traditional medicinal plant; Seoritae, and it has effects of anti-obesity, chronic cervical pain, and anti-oxidant. The present study aimed to investigate the therapeutic potential of WSY-0702 in the prevention and treatment of BPH. Several parameters including inflammatory mediators, hormones, and oxidative stress (OS) have been considered to play a role in the development of BPH. Prostate tissue damage and OS may lead to compensatory cellular proliferation with resulting hyperplastic growth. An in vitro study showed that proliferation inhibited the human prostate epithelial cell line RWPE-1 in a dose-dependent manner. In cell line, the cell cycle at the G2/M and G0/G1 phase and downregulated the expression of CyclineB1 (CCNB1) and CyclineD1 (CCND1). In addition, we measured the $H_2O_2$-induced OS damage using RWPE-1 cells. We examined the relative expression of protein involved in the regulation of prostate apoptosis: transforming growth factor (TGF)-${\beta}$, a negative growth factor able to induced prostate apoptosis under physiological conditions. These results suggest that WSY-0702 that can inhibit the growth of prostate epithelial cell by a mechanism that may involve arresting the cell cycle and downregulating CCNB1 and CCND1 expression. In addition, WSY-0702 exposure resulted in significant protective effects in $H_2O_2$-stressed PWPE-1 cells by reduction in TGF-${\beta}$ levels.

The Role of Yoga Intervention in the Treatment of Allergic Rhinitis: A Narrative Review and Proposed Model

  • Chauhan, Ripudaman Singh;Rajesh, S.K
    • CELLMED
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    • v.10 no.3
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    • pp.25.1-25.7
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    • 2020
  • Allergic Rhinitis (AR) is an IgE (immunoglobin-E) mediated inflammatory condition of upper respiratory tract; main clinical features involve runny nose, sneezing, nasal obstruction, itching and watery eyes. AR is a global problem and has large variations in incidences, currently affects up to 20% - 40% of the population worldwide. It may not be a life-threatening disease per se but indisposition from the condition can be severe and has the potential to adversely affect the daily functioning of life. Classical yoga literature indicates that, components of yoga have been used to treat numerous inflammatory conditions including upper respiratory tract. A few yoga intervention studies reported improvement in lung capacity, Nasal air flow and symptoms of allergic rhinitis. This review examined various anti-inflammatory pathways mediated through Yoga that include downregulation of pro-inflammatory cytokines and upregulation of anti-inflammatory cytokines. The hypothalaminic-pitutary-adrenal (HPA) axis and vagal efferent stimulation has been reported to mediate anti-inflammatory effect. A significant reduction is also reported in other inflammatory biomarkers like- TNF-alpha, nuclear factor kappa B (NF-κB), plasma CRP and Cortisol level. Neti, a yogic nasal cleansing technique, reported beneficial effect on AR by direct physical cleansing of thick mucus, allergens, and inflammatory mediator from nasal mucosa resulting in improved ciliary beat frequency. We do not find any study showing effect of yoga on neurogenic inflammation. In summary, Integrated Yoga Therapy may have beneficial effect in reducing symptoms and improving quality of life for patients with allergic rhinitis. Yoga may reduce inflammation through mediating neuro-endocrino-immunological network. Future studies are needed to explore the mechanism how yoga might modulate immune inflammation cascade and neurogenic inflammation at the cellular level in relevance to allergic rhinitis; the effects of kriyas (yogic cleansing techniques) also need to be evaluated in early and late phase of AR. So the proposed model could guide future research.

Expression of Ki67 and CD105 as Proliferation and Angiogenesis Markers in Salivary Gland Tumors

  • Tadbir, Azadeh Andisheh;Pardis, Soheil;Ashkavandi, Zohreh Jafari;Najvani, Ali Dehghani;Ashraf, Mohammad Javad;Taheri, Ali;Zadeh, Maryam Asad;Sardari, Yasaman
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.5155-5159
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    • 2012
  • Objective: To investigate the association between CD105 and tumor cell proliferation in salivary gland tumors. Methods: In this study, 59 samples of salivary tumors from Khalili Hospital archive, including 20 cases of pleomorphic adenoma (PA), 20 cases of mucoepidermoid carcinoma (MEC) and 19 cases of adenoid cystic carcinoma, as well as 10 cases of normal salivary gland tissue, were reviewed by immunohistochemistry (IHC) for CD105 and Ki67 staining. Results: CD105 positive vessels were absent in normal salivary gland tissue in the vicinity of tumors (51.6% of all tumors were positive). There was a statistically significant difference in frequency of CD105 staining between PA and malignant tumors and between four groups of different lesions (p<0.000) being highest in MEC. Intratumoral microvessel density was also elevated in malignant neoplasms ($2.61{\pm}3.1$) as compared to PA ($0.46{\pm}0.6$). Normal salivary glands did not express Ki67. There was a statistically significant difference in frequency and percentage of Ki67 immunoreactivity in malignant neoplasms (86.5% and $10.7{\pm}10.8$ respectively) compared to PA (50% and $0.78{\pm}0.2$) and among the four groups values were highest in MEC (p<0.000). Conclusion: n this study, it was observed a higher rate of angiogenesis and cellular proliferation was noted in malignant tumors compared to benign tumors, but no correlation was observed between these two markers.

Combined Genotype Analyses of Precursor miRNA-196a2 and -499a Variants with Hepatic and Renal Cancer Susceptibility- a Preliminary Study

  • Toraih, Eman A;Fawzy, Manal S;Elgazzaz, Mona G;Hussein, Mohammad H;Shehata, Rasha H;Daoud, Hisham G
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3369-3375
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    • 2016
  • MicroRNAs, a novel class of small non-coding RNAs, are key players in many cellular processes, including cell proliferation, differentiation, invasion and regeneration. Tissue and circulatory microRNAs could serve as useful clinical biomarkers and deregulated expression levels have been observed in various cancers. Gene variants may alter microRNA processing and maturation. Thus, we aimed to investigate the association of MIR-196a2 rs11614913 (C/T), MIR-499a rs3746444 (A/G) polymorphisms and their combination with cancer susceptibility in an Egyptian population. Sixty five renal cell carcinoma (RCC) and 60 hepatocellular carcinoma (HCC) patients and 150 controls were enrolled in the study. They were genotyped using real-time polymerase chain reaction technology. Both $miR-196a2^*T$ and $miR-499a*G$ were associated with RCC risk, but only $miR-196a^*T$ was associated with HCC development. Carriage of the homozygote combinations ($MIR196a2^*TT+MIR499a^*AA$) and ($MIR196a2^*CC+MIR499a^*GG$) was associated with 25 and 48 fold elevation of likelhood to develop RCC, respectively. The miR-196a2 SNP was also linked with larger tumor size in RCC and advanced tumor stage in HCC. miR-196a2 and miR-499a combined genotypes were associated with RCC and HCC. Further functional analysis of SNPs is required to confirm relationships between genotypes and phenotypes.